Analysing human neural stem cell ontogeny by consecutive isolation of Notch active neural progenitors

Abstract

Decoding heterogeneity of pluripotent stem cell (PSC)-derived neural progeny is fundamental for revealing the origin of diverse progenitors, for defining their lineages, and for identifying fate determinants driving transition through distinct potencies. Here we have prospectively isolated consecutively appearing PSC-derived primary progenitors based on their Notch activation state. We first isolate early neuroepithelial cells and show their broad Notch-dependent developmental and proliferative potential. Neuroepithelial cells further yield successive Notch-dependent functional primary progenitors, from early and midneurogenic radial glia and their derived basal progenitors, to gliogenic radial glia and adult-like neural progenitors, together recapitulating hallmarks of neural stem cell (NSC) ontogeny. Gene expression profiling reveals dynamic stage-specific transcriptional patterns that may link development of distinct progenitor identities through Notch activation. Our observations provide a platform for characterization and manipulation of distinct progenitor cell types amenable for developing streamlined neural lineage specification paradigms for modelling development in health and disease.

Zika Virus Disrupts Phospho-TBK1 Localization and Mitosis in Human Neuroepithelial Stem Cells and Radial Glia

“…Different early human neural stem/progenitor cell types, including neuroepithelial-like cells, have been derived from hPSCs and the developing human brain (Conti and Cattaneo, 2010; Edri et al, 2015; Elkabetz et al, 2008; Sun et al, 2008, Tailor et al, 2013). However, to the best of our knowledge, stable cell lines of neuroepithelial cells have not been derived from the dorsal forebrain (prospective NCX) or the SC.…”

“…While recent work on hPSCs has expanded the range of neural cell types attainable (Conti and Cattaneo, 2010; Edri et al, 2015; Koch et al, 2009), and a population of NES cells has been derived from the hindbrain (Tailor et al, 2013), the NES cell system we described and characterized is, to our knowledge, the first neuropotent populations derived directly from the prospective NCX and spinal cord, respectively. As such, NES cells represent a substantial advance facilitating the study of early development and neuropathology in the human CNS.…”

Evaluating cell reprogramming, differentiation and conversion technologies in neuroscience

“…During in vivo neurulation, the neural tube closes, patterning along the developmental axes takes place and the first waves of neurons are generated. In vitro , the rosette-type NPCs that can also be derived from hPSCs resemble this developmental stage 28,116,156 . During fetal and adult neurogenesis, radial glia give rise to postmitotic neurons.…”

“…The majority of iNs produced by most human iN protocols have glutamatergic properties (although some human protocols generate a minor fraction of GABAergic cells) 7,102,108,109 . As outlined above, during human hPSC differentiation, specialized NPCs are responsive to exogenous morphogens that activate lineage-specific transcription factors 28,116 . As there is no morphogen-responsive NPC intermediate in direct conversion, the co-overexpression of general pro-neuronal transcription factors with lineage-specific transcription factors has been explored (FIG.…”

A Single-Cell Roadmap of Lineage Bifurcation in Human ESC Models of Embryonic Brain Development

“…In parallel, the recent progress in modeling human brain development from pluripotent stem cells (Chambers et al, 2009; Espuny-Camacho et al, 2013; Lancaster et al, 2013; Qian et al, 2016; Shi et al, 2012) promises to supply human neural tissue at developmental stages that are typically unavailable. Although several studies have characterized differentiated cells by gene expression (Edri et al, 2015; van de Leemput et al, 2014), only one in vitro differentiation study has carried out single-cell transcriptomics (Camp et al, 2015). As these cultures presumably contain a mixture of cell types, single-cell resolution studies are essential to characterize the cell types produced in culture and to determine how they compare to primary developing tissue.…”